Broadly reactive mosaic peptide for influenza vaccine
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[0108]Chicken embryo fibroblasts (CEFs) and Mardin-Darby canine kidney (MDCK) cells were obtained from Charles River Laboratories, Inc. (Wilmington, Wash.) and the American Type Culture Collection (ATCC, Manassas, Va.), respectively. Cells were cultured in Dulbecco's Modified Eagle's Medium (DMEM) supplemented with 10% fetal bovine serum (FBS) and antibiotics. CEFs were used for propagating MVA virus. Highly pathogenic avian influenza (H5N1) virus A / Vietnam / 1203 / 04 was kindly provided by Dr. Yoshihiro Kawaoka (University of Wisconsin-Madison, Wis., USA). Highly pathogenic avian influenza (H5N1) viruses, A / Hongkong / 483 / 97, A / Mongolia / Whooper swan / 244 / 05 and A / Egypt / 1 / 08 and seasonal influenza viruses including A / Puerto Rico / 8 / 34 (PR8, H1N1) and A / Aichi / 2 / 1968 (H3N2) were kindly provided by Dr. Stacey Schultz-Chemy and Dr. Ghazi Kayali (St. Jude children's research hospital, Memphis, Term.). All viruses were propagated and titrated in MDC...
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Example II
[0134]Samples were collected for a microneutralization test at 4 weeks post-immunization. FIG. 7 shows the antibody titers for three different strains of influenza virus in mice immunized with H5M / MVA, inactivated vikeus (Baxter) or MVA alone. The antisera in immunized mice were reactive against three distinct viral clades (HPAI Clade 0, Clade 1, and Clade 2 viruses) after a single dose. The presence of neutralizing antibodies at 4 weeks indicates that the H5M vaccine provides rapid immunity and the immunity is higher than inactivated against the homologous virus. Even recent consensus approaches that have tried to control for the most diversity of input sequence have failed to simultaneously elicit immune responses against all of these clades.
TABLE 1GrpConstructsRouteN =Challenge stains1MVA-H5MID8HK / 483 / 972Inactivated H5N1SC8HK / 483 / 973MVA / LUC - controlID5HK / 483 / 974MVA-H5MID8MONG / 244 / 055Inactivated H5N1SC8MONG / 244 / 056MVA / LUC - controlID5MONG / 244 / 057MVA-H5MID8VN / 1203 / 048Ina...
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